Bioscience Methods 2025, Vol.16, No.3, 137-153 http://bioscipublisher.com/index.php/bm 151 Therefore, in specific applications, the appropriate method should be selected according to the goal. If it is a scientific research investigation, the most complete information can be obtained through whole genome sequencing/resequencing, from which mitochondrial sequences and nuclear genes can be extracted and analyzed together to obtain robust conclusions. For general species identification, COI barcode sequencing can be used first; for batch screening in customs and market supervision, specific PCR or portable detection equipment can be used to quickly lock suspicious samples. In summary, mitochondrial molecular markers provide strong technical support for abalone species identification, but we must also combine nuclear gene and morphological evidence to comprehensively evaluate the results. Different methods have their own strengths, and the accuracy and reliability of identification can be improved by complementing each other's strengths. For example, for samples with questionable identification results, 16S or nuclear gene ITS fragments can be further sequenced for verification to exclude the possibility of misjudgment of a single marker. As long as we give full play to the advantages of various methods and avoid their limitations, we can establish a robust and reliable abalone species identification system. Looking to the future, abalone systematic classification and genetic research will develop in the direction of comprehensive multi-source information and interdisciplinary cross-cutting. In terms of taxonomic research, with the acquisition of more molecular data (including mitochondrial and nuclear genomes), we have the opportunity to re-examine the systematic position and species division of the abalone genus. Perhaps hidden lineages or taxa that need to be split/merged will be discovered. To this end, future taxonomic revisions should integrate molecular phylogenetic trees, morphological and anatomical characteristics, biogeographic and ecological information, and adopt an "integrated taxonomy" approach to give a more natural species definition. At the mitochondrial genome level, in the future, it may be possible to deeply understand the genetic mechanism of abalone adaptive radiation by analyzing the selection pattern and evolutionary dynamics of whole genome sequences. For example, the study of mitochondrial gene function in species in different temperature zones is expected to reveal their energy metabolism adjustment strategies and provide a basis for the selection and improvement of abalone germplasm in response to climate change. In terms of species identification and resource protection, we expect molecular monitoring to play a greater role. Governments and scientific research institutions can establish a DNA archive of abalone germplasm resources and implement long-term molecular monitoring of wild and farmed populations. Once signs of genetic diversity decline or invasion of foreign genes are found, timely measures (such as stocking or isolation) can be taken to prevent genetic decline and germplasm contamination. At the same time, genetic technology can also be used to assist genetic selection, such as improving stress resistance traits through molecular marker-assisted selection (MAS) to achieve the cultivation of excellent varieties. From the perspective of conservation biology, more attention should be paid to the protection of abalone genetic diversity in the future. Under the pressure of global climate change and human activities, some abalone species (such as black abalone, South African abalone, etc.) face survival challenges. Molecular methods can be used to assess the genetic health of their populations and guide the establishment of conservation populations or genetic management plans with sufficient genetic diversity. For example, sequencing analysis can be used to determine the haplotype types that are preferentially retained in each conservation population to maximize the evolutionary potential of the species. In short, mitochondrial genome research will continue to be an important part of abalone systematic classification and species identification in the future, but we also need to expand our vision to the whole genome level and ecological environment level. Through multidisciplinary collaboration, we hope to fully reveal the evolutionary story of abalone and develop scientific and effective strategies to protect and utilize this precious marine biological resource. Acknowledgments We thank Mr Y. Ding from the Institute of Life Science of Jiyang College of Zhejiang A&F University for his reading and revising suggestion. Conflict of Interest Disclosure The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest.
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